| 摘要: |
| 通过对精子运动率、受精率和超微结构的观察, 研究了2003～2009 年分5 批冷冻保存的真鲷(Pagrus major)精子经1～73 个月保存后质量变化情况。冷冻精子在40 ℃水浴中解冻, 大约100～110 s转化为液态后取出, 用海水激活后观察精子运动率, 人工受精6～8 h 后观察受精率, 同时精子用2.5%戊二醛前固定, 经处理后用扫描电镜和透射电镜观察精子超微结构。结果表明, 冷冻精子的最高运动率(87.67%±2.52%)和受精率(71.33%±8.84%)都是2009 年保存1个月的精子, 保存1, 13, 26 个月的精子运动率和受精率差异不显著(P<0.05), 保存48个月后的精子运动率和受精率显著低于保存1个月的精子(P<0.05), 2003 年保存的精子(73个月)运动率(50.67%±5.31%)和受精率最低(39.56%±0.69%)。精子的超微结构损伤主要包括精子头部损伤, 线粒体损伤以及精子质膜的损伤。 |
| 关键词: 低温保存 真鲷 (Pagrus major) 精子 运动率 受精率 |
| DOI： |
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| 基金项目:中国科学院海洋研究所知识创新领域前沿项目(Y02507101Q) |
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| Quality change of Pagrus major sperm after long-term cryopreservation |
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CHEN Ya-kun,LIU Qing-hua,ZHAO Chun-yan,XIAO Zhi-zhong,XU Shi-hong,Ma Dao-yuan,LI Jun
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| Abstract: |
| After cryopreservation (1-73 months), the quality of Pagrus major sperm, which was cryopreserved from 2003 to 2009 in 5 batches, was examined by observing motility, fertilization rate and ultrastructures. The cryopreserved sperm were thawed in 40 oC water bath for 100~110 s. After activation with seawater, the motility of the sperm was estimated; 6-8 h after artificial fertilization, fertilization rate was examined; and after being fixed with by 2.5% glutaraldehyde, the ultrastructure of sperm was studied with scanning electron microscope and transmission electron microscope. The highest motility (87.67%±2.52%) and fertilization rate (71.33%± 8.84%) were obtained in sperm after a 1-month cryopreservation, even though they were not significantly different from those of the sperm after a 13-month or and 26-month cryopreservation (P>0.05). After a 48-month cryopreservation, the motility and fertilization rate decreased significantly in comparison with those of sperm after a 1-month cryopreservation (P<0.05). The lowest motility (50.67% ± 5.31%) and fertilization rate (39.56% ± 0.69%) were obtained in sperm cryopreserved for 73 months; main ultrastructural damage was observed in the head, mitochondria and membranes of sperm. |
| Key words: cryopreservation Pagrus major sperm motility fertilization rate |
